Se cell line nucleofector x kit

HEK293T cells were seeded at 1.25 × 10⁴ cells per well into 96-well flat bottom cell culture plates (Corning) for DNA on-target experiments or at 6.25 × 10⁴ cells per well into 24-well cell culture plates (Corning) for DNA off-target experiments. 24 hours post-seeding, cells were transfected with 30 ng of control or base/prime editor plasmid and 10 ng of peg or gRNA plasmid (and 3.3 ng nicking gRNA plasmid for PE3) using 0.3 μL of TransIT-X2 (Mirus) lipofection reagent for experiments in 96-well plates, or 150 ng control or base editor plasmid and 50 ng gRNA, and 1.5 μL TransIT-X2 for experiments in 24-well plates. K562 cells were electroporated using the SF Cell Line Nucleofector X Kit (Lonza), according to the manufacturer’s protocol with 2 × 10⁵ cells per nucleofection and 800 ng control or base/prime editor plasmid, 200 ng gRNA or pegRNA plasmid, and 83 ng nicking gRNA plasmid (for PE3). U2OS cells were electroporated using the SE Cell Line Nucleofector X Kit (Lonza) with 2 × 10⁵ cells and 800 ng control or base/prime editor plasmid, 200 ng gRNA or pegRNA, and 83 ng nicking gRNA (for PE3). HeLa cells were electroporated using the SE Cell Line 4D-Nucleofector X Kit (Lonza) with 5 × 10⁵ cells and 800 ng control or base/prime editor, 200 ng gRNA or pegRNA, and 83 ng nicking gRNA (for PE3). 72 hours post-transfection, cells were lysed for extraction of genomic DNA (gDNA).